A topoisomerase II-dependent G2 cycle checkpoint in mammalian cells

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Abstract
THE enzyme DNA topoisomerase II, which removes the caten-ations formed between the DNA molecules of sister chromatids during replication1 and is a structural component of chromosome cores2 , is needed for chromosome condensation in yeast3 and in Xenopus extracts4–6. Inhibitors of topoisomerase II arrest mam-malian cells before mitosis in the G2 phase of the cell cycle7, but also produce DNA damage, which causes arrest through established checkpoint controls8. It is open to question whether cells need topoisomerase II to leave G2, or control late-cycle progres-sion in response to its activity. Bisdioxopiperazines are topoisomerase II inhibitors that act without producing direct DNA damage9; the most potent, ICRF-193, blocks mammalian entry into but not exit from mitosis. Here we show that checkpoint-evading agents such as caffeine override this block to produce abortively condensed chromosomes, indicating that topoisomerase II is needed for complete condensation. We find that exit from G2 is regulated by a catenation-sensitive checkpoint mechanism which is distinct from the G2-damage checkpoint.